Part:BBa_K5136037
mt3
Biology
MT3
The metallothioneins (MTs) are a class of low molecular weight and cysteine-rich metal binding proteins, and each one of them can bind to 6-9 heavy metal ions. The MTs are expressed as intracellular protein and are primarily responsible for metal regulation in cells of living organisms. General MTs can widely non-covalently bind divalent heavy metal ions, such as Zn2+,Ni2+, Pb2+, Hg2+, Cd2+, as well as As3+. MT3 has a excellent adsorption effect on ordinary metal ions. (1).
Usage and Design
To absorb Cd2+ in waste water, we introduced MntA membrane protein in E. coli, which translocates external Cd2+ into the cell. To increase the strain's tolerance to Cd2+, we produce intracellular MTs in engineered bacteria, mitigating the cell damage of Cd2+. This basic part (BBa_K5136037) which codes the MT3 was constructed and then used for the construction of the composite part (BBa_K5136230) .
Characterization
Agarose gel electrophoresis (AGE)
I0500 promoter was employed to start the expression of MT3 (BBa_K5136037) in E. coli BL21(DE3). The basic part (BBa_K5136037) is a component of the composite part (BBa_K5136230). The composite part (BBa_K5136230) constructed was introduced into the backbone plasmid (pSB1C3) through standard assembly and transformed into E. coli BL21(DE3).The positive clones were selected, and colony PCR and gene sequencing were used to verify that the clones were correct . Target bands (2106 bp) can be observed at the position around 2000bp (Figure 1).
Reference
1. A. A. Uçkun, M. Uçkun, S. Akkurt, Efficiency of Escherichia Coli Jm109 and Genetical Engineering Strains (E. Coli MT2, E. Coli MT3) in Cadmium Removal from Aqueous Solutions. Environ. Technol. Innovation 24, 12 (2021). Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 3
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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